Home > Article: cGMP Chromatography Columns & Custom Packing
cGMP Chromatography Columns & Custom Packing
Post polishing filtration, the downstream purification steps involve chromatographic separation and purification of the target protein(s). In cGMP environments it is essential that effective column packing can be demonstrated for the process to perform as designed. In-house column packing can be challenging as this is not an everyday procedure. A non-optimally packed column will yield poor and uneven flow and poor resolution due to band broadening. This has led to the adaption of prepacked and single use or multi-cycle limited use Chromatography columns being used in critical processes.
Pure Process through its sister production sites in the UK provides a range of pre-packed, scalable, single-use Chromatography columns in a range of sizes to meet the industry needs. We can supply the customers’ specific columns with their choice of resin, pre-packed and tested, ready to use thus saving time, resources, and process performance.
GMP Bioprocessing columns are pre-packed and ready to use with a range of standard resins (see below) or resin for affinity purification of native and recombinant adeno-associated virus (AAV) subtypes. The AAVX resin is the gold standard resin used for the purification of AAV particles.
Our pre-packed cGMP Bioprocessing scale columns offer high specificity, capacity and maximise yield by reducing process steps.
All columns are packed in a controlled non-classified ISO Class 7 clean room and procedures at our UK production sites to Quality Standards - ISO 9001 QMS
All direct fluid contact materials meet USP Class VI and USP <88> requirements for In Vivo Biological Reactivity
All fluid contact components are free from materials of animal origin and compliant with EMEA 410/01
QC Lot Release Testing:
Theoretical plate numbers (HETP)
Peak asymmetry
Bioburden (<10 CFU / 100ml)
Endotoxin level (<0.25 EU/ml)
Flow / pressure measurement (on request)
Why use our Column packing services?
With our Comprehensive Large Scale Custom Column Packing Service (cGMP/non-cGMP grade), Pure Process becomes your resin packing service for disposable and scalable chromatography columns providing a service that is Cost-effective with fast turn-around times and availability.
Typically, Non-GMP columns can be packed and tested ready for release in 2 weeks, and GMP columns can be ready in 5-6 weeks. Once a pattern is established, we can of course set up a buffer stock and supply in a just in time (JIT) basis – acting in true partnership with the client.
Our columns are compatible with all preparative resins above 20 µm bead size e.g. IEX, HIC, Protein A, Heparin etc.
As a reliable partner we aim to supply columns that are industry standard and compatible with your existing chromatography equipment hence our columns are rated up to 5 bar pressure rating.
We can perform Shipping tests to ensure the tested columns arrive on site ready to use. Additionally, our proprietary lab and bioprocess columns significantly reduce validation time and costs.
Our high-quality columns and custom column packing service uses low cost, high quality disposable plastic columns making outsourcing this service to Pure Process a viable and cost efficient alternative.
So, what resins do we offer as standard and what can we pack?
Chromatography Resins
We supply a range of resins manufactured within our group companies in conjunction with the column packing service above. Chemistries like:
Hydrophobic Interaction Chromatography (HIC) Resins
HIC is a versatile technique and can show high selectivity to individual molecules according to their exposed hydrophobic zones. It is particularly useful for intermediate and final-stage purifications and are specially designed for the purification of biological molecules based on their hydrophobicity profiles.
HIC media requires much milder purification conditions than reversed phase chromatography (RPC) media and biological activity can be maintained in HIC separations unlike RPC operations.
HIC media normally binds at moderate to high salt concentrations and is therefore ideally placed step after an IEX step where molecules are usually eluted at high salt conditions.
SepFast™ HIC media offers a broad range of choices from C4 to C8. In general, the longer the carbon chain, the higher the surface hydrophobicity.
Features of our HIC resins:
High sample loading capacity and High separation power
Chemically stable Agarose base matrix
Can be produced in high and low ligand densities, alternate crosslinking levels and different bead sizes to optimise the media to the separation
Available cGMP grade for full manufacturing scale
|
Bead Size and Type |
Typical Application |
|
4HF serial (50µm – 150µm) |
Purification of most medium to large proteins |
|
6HF serial (50µm – 150µm) |
Purification of peptides or smaller proteins |
|
Large Beads (150µm – 350µm) |
Purification of proteins from crude or viscous samples |
|
HighRes (25µm – 50µm) |
Purification of proteins that require high resolution |
Super Loose Resins
The mean bead size range is 35 μm. High selective binding capacity is the key benefit of our Super resins.
-
Chemistries: Ni-NTA, Co-NTA, Glu, Protein A, Protein G and Heparin
Fastback Loose Resins
The mean bead size of our Fastback resins range is 90 μm, ideal for fast purification of target proteins.
Chemistries: Protein A & Protein G, Co IMAC, Ni IMAC
Ion-Exchange
SepFast DUO™ IEX is a class of unique ion-exchange resins having size-exclusion inert shell and ion-exchanger core. This dual functionality dramatically boosts the selectivity and purification power based on molecular weights and electrostatic charges.
SepFast Supor Ion-Exchange
Our Novel SepFast Supor chromatography media has a combination of very large pores (accessible by viruses) and small pores (accessible to proteins). It has high binding capacity to both viral particles and proteins almost independent of flow velocity.
ViralPolish®
ViralPolish® chromatography media is specifically designed for flow-through virus purification use. It comprises dual layer agarose beads with an inert external shell with tightly controlled pore size and internal multi-functional ligands for rapid high-capacity binding of impurities.
Viralpolish® media is ideal for rapid clean-up, either as a first step or as a second step, to purify virus particles. Large particles, such as viruses, are excluded from the beads which can be packed in a column (our custom column packing service) where the virus will pass through the column bed and be collected in the flow-through fraction, whilst impurities will be captured within the beads. This affords a very gentle purification process resulting in efficient clean up and high recovery of active virus particles.
The beads can also be used in batch mode; when added to a flask of virus containing media the beads will selectively allow small molecules to enter and be retained, whilst the virus is excluded and remains in the external media. Our Single-Use Filtration technology can be used to capture and separate the beads after the process step (see: Single Use Filtration).
A typical process purification strategy for virus would be to use ViralPolish® as an initial step, to clean up the sample, prior to binding and elution, typically from an anion exchange media designed for virus purification such as SepFast Supor Q.
Alternatively, very dilute samples can be purified by first passing through a SepFast Supor Q anion exchange column to concentrate the sample followed by passing through ViralPolish column to remove residual impurities.
ViralPolish® is available in three different outer shell pore sizes, and two different impurity binding chemistries.
AAVX Resin
Specifically designed resin for the large scale Purification of Adeno-associated Viruses (AAV) for Gene Therapy Applications which is supplied exclusively pre-packed in our cGMP custom columns specifically for the purification of all AAV serotypes (AAV1-AAV9).
Evaluation / Testing
If you need laboratory scale products for evaluation for any of the above please contact us or visit our sister websites: www.labunlimited.com or UK: www.labunlimited.co.uk
Please contact us for further information:
Sailesh Patel
Phone: 00353 1 4523432
E-mail: info@pureprocess.eu
Website: www.pureprocess.eu